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Single-cell proteomic chip for profiling intracellular signaling pathways in single tumor cells

Shi, Qihui and Qin, Lidong and Wei, Wei and Geng, Feng and Fan, Rong and Shin, Young Shik and Guo, Deliang and Hood, Leroy and Mischel, Paul S. and Heath, James R. (2012) Single-cell proteomic chip for profiling intracellular signaling pathways in single tumor cells. Proceedings of the National Academy of Sciences of the United States of America, 109 (2). pp. 419-424. ISSN 0027-8424. PMCID PMC3258586. https://resolver.caltech.edu/CaltechAUTHORS:20120203-103706722

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Abstract

We describe a microchip designed to quantify the levels of a dozen cytoplasmic and membrane proteins from single cells. We use the platform to assess protein–protein interactions associated with the EGF-receptor-mediated PI3K signaling pathway. Single-cell sensitivity is achieved by isolating a defined number of cells (n = 0–5) in 2 nL volume chambers, each of which is patterned with two copies of a miniature antibody array. The cells are lysed on-chip, and the levels of released proteins are assayed using the antibody arrays. We investigate three isogenic cell lines representing the cancer glioblastoma multiforme, at the basal level, under EGF stimulation, and under erlotinib inhibition plus EGF stimulation. The measured protein abundances are consistent with previous work, and single-cell analysis uniquely reveals single-cell heterogeneity, and different types and strengths of protein–protein interactions. This platform helps provide a comprehensive picture of altered signal transduction networks in tumor cells and provides insight into the effect of targeted therapies on protein signaling networks.


Item Type:Article
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1073/pnas.1110865109DOIArticle
http://www.pnas.org/content/109/2/419.abstractPublisherArticle
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3258586/PubMed CentralArticle
ORCID:
AuthorORCID
Hood, Leroy0000-0001-7158-3678
Heath, James R.0000-0001-5356-4385
Additional Information:© 2012 by the National Academy of Sciences. Edited by Chad A. Mirkin, Northwestern University, Evanston, IL, and approved November 17, 2011 (received for review July 6, 2011). Published online before print December 27, 2011. We thank Bruz Marzolf and Pamela Troisch for printing DNA microarrays, and the University of California, Los Angeles nanolab for photomask fabrication. This work was funded by the National Cancer Institute Grant 5U54 CA119347 (to J.R.H., principal investigator), The Ben and Catherine Ivy Foundation, the Goldhirsch Foundation, and the Grand Duchy of Luxembourg. Author contributions: Q.S., L.Q., and J.R.H. designed research; Q.S., L.Q., F.G., R.F., Y.S.S., and D.G. performed research; Q.S., W.W., L.H., P.S.M., and J.R.H. analyzed data; and Q.S., W.W., L.H., P.S.M., and J.R.H. wrote the paper. The authors declare no conflict of interest.
Funders:
Funding AgencyGrant Number
National Cancer Institute5U54 CA119347
Ben and Catherine Ivy FoundationUNSPECIFIED
Goldhirsh Foundation UNSPECIFIED
Grand Duchy of Luxembourg UNSPECIFIED
Issue or Number:2
PubMed Central ID:PMC3258586
Record Number:CaltechAUTHORS:20120203-103706722
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20120203-103706722
Official Citation:Qihui Shi, Lidong Qin, Wei Wei, Feng Geng, Rong Fan, Young Shik Shin, Deliang Guo, Leroy Hood, Paul S. Mischel, and James R. Heath Single-cell proteomic chip for profiling intracellular signaling pathways in single tumor cells PNAS 2012 109 (2) 419-424; published ahead of print December 27, 2011, doi:10.1073/pnas.1110865109
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:29122
Collection:CaltechAUTHORS
Deposited By: Ruth Sustaita
Deposited On:03 Feb 2012 18:56
Last Modified:03 Oct 2019 03:38

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